Anti rad6

Proteins were separated by standard 10 - 15% SDS-PAGE loaded in Laemmli buffer. Samples were transferred to PVDF membrane (ThermoFisher), and immunoblotting was performed using the following antibodies: anti-K63 ubiquitin (1:4,000; EMD Millipore, cat. no. 05-1308, clone apu3), anti-actin (1:6,000; Cell Signaling, cat. no. 4967), anti-GAPDH (1:4,000; Abcam, cat. no. ab9485), anti-Myc (1:5,000; ThermoFisher, cat. no. R950-25 and PA1-981), anti-Rad6 (1:8,000; Abcam, cat. no. ab31917), anti-Rps3/uS3 (1:6,000; Cell Signaling, cat. no. 9538S), anti-Rpl11/uL5 (1:6,000; Cell Signaling, cat. no. 18163S), anti-HA (1:10,000; ThermoFisher, cat. no. 71-5500), anti-Rps20/uS10 (1:9,000; ThermoFisher, cat. no. PA5-75383), anti-ubiquitin (1:10,000; Cell Signaling Technology cat. No. 3936S), anti-FLAG (1:3,000; MilliporeSigma, cat. no. F3165), anti-puromycin (1:4,000; MilliporeSigma, cat. no. MABE343), anti-ubc13 (1:1,000; Novus Biologicals, cat. no. NBP1-76593). Anti-mouse IgG (1:8,000-12,000; Cytiva, ca. no. NA931), anti-rabbit IgG (1:6,000-12,000; Cytiva, cat. no. NA934), and Anti-mouse IgG2a (1:10,000-1:12,000; Abcam, cat. no. ab97245) secondary antibodies conjugated with HRP and ECL Prime detection reagents were acquired from Cytiva. All antibodies have been validated by the manufacturer or are expected to react with the species used in this study based on sequence similarity.